Lambda DNA mini-preps?
palomb at beagle.Colorado.EDU
Sat Dec 4 22:06:38 EST 1993
rocket1 at bu.edu (Richard Near) writes:
>If someone has a good method to prepare lambda DNA as minipreps, I'd
>be interested. I've been growing 50ml (using LE392), doing a PEG
>precip., proteinase K, Phenol/Chloroform technique. I've been getting
>low yields, but the DNA is at least cuttable.
> What can I do to increase my yields (I get good lysis) yet keep the
>prep simple. The DNA should be clean enough for clone mapping and
>I intend to use it as a template for PCR sequencing.
Try the method of Santos (NAR 19:5442;1991). He uses ZnCl2
to precipitate out phage particles and a simple lysis buffer to get the DNA
Personally, I find I still need a phenol step, but that's probably
becuase I do my preps from plates (with agarose) rather than liquid more
often than not. I usually get around 10 micrograms from a 150 mm plate,
more than enough for most diagnostic purposes.
As long as I'm here, anyone know the chemistry involved in using
ZnCl2 to precipitate?
HA! ...*Pronoun trouble.* It's not, "He doesn't have to shoot *you*
now," it's, "He doesn't have to shoot *me* now." Well, I say he DOES
have to shoot me now! So SHOOT ME NOW!!  palomb at beagle.colorado.edu
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